Deficiency of pRb Decreases Hepatocyte Sensitivity to TGF-? / 中国生物工程杂志

ABSTRACT

Aim:to detect how Rb-deficiency will affect responses to TGF-? induced cell cycle arrest of hepatocyte.Methods:primary hepatocytes were isolated and cultured,and Rb-specific adenovirus siRNA was transformed into cells.Then TGF-? was added daily and cell growth and cell cycles variations.Western blot and FQ-RT-PCR were adopted to detect pRb,E2F,c-MYC and p16 expression changes.Results:Primary hepatocytes were isolated and infected with Rb-specific siRNA adenovirus.In control cells,treatment with TGF-? prevented cells to enter S phase via decreased cMYC activity,activation of P16INK4A activity.In Rb-null hepatocytes,E2F and cMYC activity decreased slightly but P16INK4A was not activated and the great majority of cells continued cycling.Rb is therefore central to TGF?-induced cell cycle arrest in hepatocytes.Conclusion:The present results show that otherwise genetically normal hepatocytes with disabled Rb genes respond less well to the antiproliferative effects of TGF-?.